title: Isolation of murine postnatal brain microglia for phenotypic characterization using magnetic cell separation technology.
abstract: To shorten the time between brain harvesting and microglia isolation, and characterization, we utilized the MACS(®) neural dissociation kit followed by OctoMACS(®) CD11b magnetic bead isolation technique to positively select for brain microglia expressing the pan-microglial marker CD11b, a key subunit of the membrane attack complex (MAC). This protocol yields a viable and highly pure (>95%) microglial population of approximately 500,000 cells per pup that is amenable for in vitro characterization within hours or days after being harvested from brain tissue. Primary microglia from C57Bl/6 mice were plated for next-day analyses of morphology and cellular markers by immunocytochemistry or for analysis of gene expression under resting or LPS-stimulated conditions. The ease of isolation enables investigators to perform molecular and cellular analyses without having to wait 1-2 weeks to isolate microglia by conventional methods involving mechanical agitation to dislodge these from astrocyte beds.
authors.affiliation: Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Ashley S
authors.initials: AS
authors.lastname: Harms
authors.forename: Malú G
authors.initials: MG
authors.lastname: Tansey
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology*; D002469:Cell Separation / Q000379:methods*; D051379:Mice; D017628:Microglia / Q000166:cytology*